Phenethylamine

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

1999

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)

GLP compliance:

yes (incl. QA statement)

Remarks:

testing lab.

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Phenethylamine

EC Number:

200-574-4

EC Name:

Phenethylamine

Cas Number:

64-04-0

Molecular formula:

C8H11N

IUPAC Name:

2-phenylethanamine

Test material form:

liquid

Details on test material:

Name of the test substance used in the study report: 2-Phenylethylamin
Purity: 99 .7 area % (GC)

Method

Target gene:

S. typhimurium and E. coli

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

Aroclor-induced rat liver S-9 mix

Test concentrations with justification for top dose:

20 ug - 5000 ug/plate (standard plate test and preincubation test)

Vehicle / solvent:

Due to the limited insolubility of the test substance in water, DMSO was selected as the vehicle, which had been demonstrated to be suitable in bacterial reverse mutation tests and for which historical control data are available.

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

other: with metabolic activation: 2-aminoanthracene; without metabolic activation: N-methyl-N'-nitro-N-nitrosoquanidine, 4-nitro-o-phenylendiamine, 9-aminoacridine and N-ethyl-N'-nitro-N-nitrosoquanidine

Details on test system and experimental conditions:

Standard plate test
Salmonella typhimurium:
Test tubes containing 2-ml portions of soft agar (overlay agar), which consists of 100 ml agar (0 .6 % agar + 0 .6% NaCl) and 10 ml amino acid solution (minimal amino acid solution for the determination of mutants : 0 .5 mM histidine + 0 .5 mM biotin) are kept in a water bath at 45°C, and the remaining components are added in the following order :
0.1 ml test solution or vehicle; 0.1 ml fresh bacterial culture; 0.5 ml S-9 mix (in tests with metabolic activation) or 0.5 ml phosohate buffer (in tests without metabolic activation)
After mixing, the samples are poured onto Vogel-Bonner agar plates (minimal glucose agar plates) within approx. 30 seconds.
Composition of the minimal glucose agar : 980 ml aqua dest.; 20 ml Vogel-Bonner E medium; 15 g Difco bacto agar; 20 g D-glucose, monohydrate. After incubation at 37°C for 48 - 72 hours in the
dark, the bacterial colonies (his* revertants) are counted.
Escherichia coli:
Test tubes containing 2-ml portions of soft agar (overlay agar), which consists of 100 ml agar (0 .6% agar + 0 .6% NaCl) and 10 ml amino acid solution (minimal amino acid solution for the determination of mutants : 0 .5 mM tryptophan) are kept in a water bath at 45°C, and the remaining components are added in the following order : 0 .1 ml test solution or vehicle; 0 .1 ml fresh bacterial cultur e
0 .5 ml S-9 mix (in tests with metabolic activation) or 0 .5 ml phosphate buffer (in tests without metabolic activation). After mixing, the samples are poured onto minimal agar plates within approx . 30 seconds.
The composition of the minimal agar (SAl selective agar) is based on the description of Green, M .H .L . and Muriel, W .J . (5), with the exception of solution E (tryptophan solution), which has previously been added to the soft agar : 300 ml solution B (agar ); 100 ml solution A (saline solution); 8 ml solution C (glucose solution); 10 ml solution D (casein solution). After incubation at 37°C for 48 - 72 hours in the dark, the bacterial colonies (trp* revertants) are counted.

Preincubation test
0 .1 ml test solution or vehicle, 0 .1 ml bacterial suspension and 0 .5 ml S-9 mix are incubated at 37°C for the duration of about 20 minutes using a shaker. Subsequently, 2 ml of soft agar is added and, after mixing, the samples are poured onto the agar plates within approx. 30 seconds. After incubation at 37°C for 48 - 72 hours in the dark, the bacterial colonies are counted.

Evaluation criteria:

The test chemical is considered positive in this assay if the following criteria are met :
• A dose-related and reproducible increase in the number of revertant colonies, i .e . about doubling of the spontaneous mutation rate in at least one tester strain either without S-9 mix or after adding a metabolizing system.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

other: A bacteriotoxic effect was observed in the standard plate test from about 2500 ug/plate onward.

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

other: A bacteriotoxic effect was observed in the standard plate test from about 2500 ug/plate onward.

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

An increase in the number of his+ or trp+ revertants was not observed in the standard plate test or in the preincubation test either without S-9 mix or after the addition of a metabolizing system.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

According to the results of the present study, the test substance is not mutagenic in the Salmonella typhimurium/Escherichia coli reverse mutation assay under the experimental conditions chosen here.